Method for producing a biological tissue construct and use of specifically obtained autologous cells

Abstract

The method for preparing a tissue construct for medical purposes uses endothelial progenitor cells (EPC) which have not been passaged multiple times and have a content of EOEC (early outgrowth endothelial progenitor cells) and LOEC (late outgrowth endothelial progenitor cells). These cells and fibroblasts and/or muscle cells, viz. myoblasts, myofibroblasts, smooth muscle cells or the progenitors thereof, are, in the form of living cells, seeded onto a matrix or introduced into a matrix in order to yield the tissue construct following further treatment steps. The matrix is preferably a protein preparation, more particularly a fibrinogen preparation. Both the cells and the fibrinogen preparation can be obtained from a single blood sample from a patient. Apart from the preparation of bypass materials, prosthetic vascular graft, tissue patches, conduits and the like, the EOEC-containing EPC culture or suspension is suitable as means for cell transplantation.

The invention claimed is: 
     
       1. A method of making a tissue construct for medical purposes, comprising the steps of:
 a) seeding onto or into a biocompatible matrix endothelial progenitor cells (EPC) having a content of both early outgrowth endothelial progenitor cells (EOEC) and late outgrowth endothelial progenitor cells (LOEC), wherein the endothelial progenitor cells have not been passaged multiple times, and wherein the LOEC's are CD133 + , and 
 b) simultaneously or sequentially seeding into or onto the biocompatible matrix one or more cell types selected from the group consisting of myoblasts, myofibroblasts, smooth muscle cells, smooth muscle progenitor cells and fibroblasts. 
 
     
     
       2. The method according to  claim 1 , wherein at least the endothelial progenitor cells are obtained from blood. 
     
     
       3. The method according to  claim 1 , wherein the endothelial progenitor cells are obtained from adipose tissue. 
     
     
       4. The method according to  claim 1 , wherein the matrix is composed of at least one material selected from the group consisting of an in vivo nondegradable biocompatible material, an in vivo degradable biodegradable synthetic material, and an in vivo degradable biodegradable biological material. 
     
     
       5. The method according to  claim 4 , wherein the cells are introduced into the matrix and the matrix is a protein preparation which is initially in a liquid, pasty or gel-like state and is subsequently stabilized. 
     
     
       6. The method according to  claim 5 , wherein the protein preparation is a fibrinogen preparation obtained from blood. 
     
     
       7. The method according to  claim 5 , wherein the protein preparation containing the cells is initially in a liquid and is subsequently stabilized and is cast onto a plate or introduced into a mold, allowed to set, and, thereafter, demolded to yield a dimensionally stable construct. 
     
     
       8. The method according to  claim 7 , wherein the dimensionally stable construct, after demolding from the mold, is perfused in a bioreactor. 
     
     
       9. The method of  claim 4 , wherein said matrix is a prosthesis. 
     
     
       10. The method of  claim 1 , wherein said early outgrowth endothelial progenitor cells and said late outgrowth endothelial progenitor cells are not passaged in cell culture. 
     
     
       11. A method of making a tissue construct for medical purposes, wherein the tissue construct is a patient-specific bioartificial tissue construct and wherein the tissue construct is colonized with living cells in vitro, comprising the steps of:
 a) seeding two types of living cells into or onto a biocompatible matrix wherein the cell types are early outgrowth endothelial progenitor cells (EOEC) and late outgrowth endothelial progenitor cells (LOEC) which are obtained from the blood of said patient and which are obtained without multiple passaging, and wherein the LOEC's are CD133 + , and 
 b) simultaneously or sequentially seeding into or onto the biocompatible matrix one or more cell types selected from the group consisting of myoblasts, myofibroblasts, smooth muscle cells, smooth muscle progenitor cells, and fibroblasts. 
 
     
     
       12. The method according to  claim 11 , wherein the tissue construct is a bypass material, a prosthetic vascular graft, a tissue patch, a tissue replacement part or a conduit. 
     
     
       13. The method according to  claim 11 , wherein additional cell types comprising autologous cells from the construct recipient are seeded into or onto the biocompatible matrix. 
     
     
       14. The method of  claim 11 , wherein said early outgrowth endothelial progenitor cells and said late outgrowth endothelial progenitor cells are not passaged in cell culture. 
     
     
       15. A method of making a tissue construct for medical purposes, comprising the steps of:
 a) seeding onto or into a biocompatible matrix endothelial progenitor cells (EPC) having a content of both early outgrowth endothelial progenitor cells (EOEC) and late outgrowth endothelial progenitor cells (LOEC), wherein the endothelial progenitor cells have not been passaged multiple times, and 
 b) simultaneously or sequentially seeding into or onto the biocompatible matrix one or more cell types selected from the group consisting of myoblasts, myofibroblasts, smooth muscle cells, smooth muscle progenitor cells and fibroblasts, 
 wherein said early outgrowth endothelial progenitor cells and said late outgrowth endothelial progenitor cells are passaged once in cell culture. 
 
     
     
       16. The method according to  claim 15 , wherein at least the endothelial progenitor cells are obtained from blood. 
     
     
       17. The method according to  claim 15 , wherein the endothelial progenitor cells are obtained from adipose tissue. 
     
     
       18. The method according to  claim 15 , wherein the matrix is composed of at least one material selected from the group consisting of an in vivo nondegradable biocompatible material, an in vivo degradable biodegradable synthetic material; and an in vivo degradable biodegradable biological material. 
     
     
       19. The method according to  claim 18 , wherein the cells are introduced into the matrix and the matrix is a protein preparation which is initially in a liquid, pasty or gel-like state and is subsequently stabilized. 
     
     
       20. The method according to  claim 19 , wherein the protein preparation is a fibrinogen preparation obtained from blood. 
     
     
       21. The method according to  claim 20 , wherein the protein preparation containing the cells is cast onto a plate or introduced into a mold, allowed to set, and, thereafter, demolded to yield a dimensionally stable construct. 
     
     
       22. The method according to  claim 21 , wherein the dimensionally stable construct, after demolding from the mold, is perfused in a bioreactor. 
     
     
       23. A method of making a tissue construct for medical purposes, wherein the tissue construct is a patient-specific bioartificial tissue construct and wherein the tissue construct is colonized with living cells in vitro, comprising the steps of:
 a) seeding two types of living cells into or onto a biocompatible matrix wherein the cell types are early outgrowth endothelial progenitor cells (EOEC) and late outgrowth endothelial progenitor cells (LOEC) which are obtained from the blood of said patient and which are obtained without multiple passaging, and 
 b) simultaneously or sequentially seeding into or onto the biocompatible matrix one or more cell types selected from the group consisting of myoblasts, myofibroblasts, smooth muscle cells, smooth muscle progenitor cells, and fibroblasts, 
 wherein said early outgrowth endothelial progenitor cells and said late outgrowth endothelial progenitor cells are passaged once in cell culture. 
 
     
     
       24. The method according to  claim 23 , wherein the tissue construct is a bypass material, a prosthetic vascular graft, a tissue patch, a tissue replacement part or a conduit. 
     
     
       25. The method according to  claim 23 , wherein additional cell types comprising autologous cells from the construct recipient are seeded into or onto the biocompatible matrix. 
     
     
       26. A method of making a tissue construct for medical purposes, comprising the steps of:
 a) seeding onto or into a biocompatible matrix endothelial progenitor cells (EPC) having a content of both early outgrowth endothelial progenitor cells (EOEC) and late outgrowth endothelial progenitor cells (LOEC), wherein the endothelial progenitor cells have not been passaged multiple times, and 
 b) simultaneously or sequentially seeding into or onto the biocompatible matrix one or more cell types selected from the group consisting of myoblasts, myofibroblasts, smooth muscle cells, smooth muscle progenitor cells and fibroblasts, wherein the matrix is composed of at least one material selected from the group consisting of an in vivo nondegradable biocompatible material, an in vivo degradable biodegradable synthetic material, and an in vivo degradable biodegradable biological material, wherein the cells are introduced into the matrix and the matrix is a protein preparation which is initially in a liquid or pasty state and is subsequently stabilized, and wherein the protein preparation containing the cells is cast onto a plate or introduced into a mold, allowed to set, and, thereafter, demolded to yield a dimensionally stable construct. 
 
     
     
       27. The method according to  claim 26 , wherein the dimensionally stable construct, after demolding from the mold, is perfused in a bioreactor. 
     
     
       28. The method according to  claim 26 , wherein at least the endothelial progenitor cells are obtained from blood. 
     
     
       29. The method according to  claim 26 , wherein the endothelial progenitor cells are obtained from adipose tissue. 
     
     
       30. The method of  claim 26 , wherein said matrix is a prosthesis. 
     
     
       31. The method of  claim 26 , wherein said early outgrowth endothelial progenitor cells and said late outgrowth endothelial progenitor cells are not passaged in cell culture. 
     
     
       32. The method of  claim 26 , wherein the protein preparation is a fibrinogen preparation obtained from blood.